<scp>RNAi</scp>‐mediated silencing of Mediterranean fruit fly (<scp><i>Ceratitis capitata</i></scp>) endogenous genes using orally‐supplied double‐stranded <scp>RNAs</scp> produced in <scp><i>Escherichia coli</i></scp>

Ortolá, Beltrán; Urbaneja, Alberto; Eiras, Marcelo; Pérez‐Hedo, Meritxell; Daròs, José‐Antonio

doi:10.1002/ps.7839

Cited by 4 publications

(1 citation statement)

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“…To produce dsRNAs, the incorporation of a self-processing group-I intron cDNA between the inverted repeat is needed to stabilize the expression plasmids, while the intron RNA is efficiently excised from the final chimera, contributing to its compaction. An additional sequence of the intron in a permuted configuration flanking the inverted repeat allows the production of circular dsRNAs without the viroid scaffold [ 292 , 293 ]. This system is also interesting as some viroids such as ELVd are the only known pathogens able to infect the chloroplast.…”

Section: Sources Of Dsrnas With Insecticidal Effectmentioning

confidence: 99%

RNA Interference in Insects: From a Natural Mechanism of Gene Expression Regulation to a Biotechnological Crop Protection Promise

Ortolá,

Daròs

2024

Biology

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Insect pests rank among the major limiting factors in agricultural production worldwide. In addition to direct effect on crops, some phytophagous insects are efficient vectors for plant disease transmission. Large amounts of conventional insecticides are required to secure food production worldwide, with a high impact on the economy and environment, particularly when beneficial insects are also affected by chemicals that frequently lack the desired specificity. RNA interference (RNAi) is a natural mechanism gene expression regulation and protection against exogenous and endogenous genetic elements present in most eukaryotes, including insects. Molecules of double-stranded RNA (dsRNA) or highly structured RNA are the substrates of cellular enzymes to produce several types of small RNAs (sRNAs), which play a crucial role in targeting sequences for transcriptional or post-transcriptional gene silencing. The relatively simple rules that underlie RNAi regulation, mainly based in Watson–Crick complementarity, have facilitated biotechnological applications based on these cellular mechanisms. This includes the promise of using engineered dsRNA molecules, either endogenously produced in crop plants or exogenously synthesized and applied onto crops, as a new generation of highly specific, sustainable, and environmentally friendly insecticides. Fueled on this expectation, this article reviews current knowledge about the RNAi pathways in insects, and some other applied questions such as production and delivery of recombinant RNA, which are critical to establish RNAi as a reliable technology for insect control in crop plants.

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Section: Sources Of Dsrnas With Insecticidal Effectmentioning

confidence: 99%

RNA Interference in Insects: From a Natural Mechanism of Gene Expression Regulation to a Biotechnological Crop Protection Promise

Ortolá,

Daròs

2024

Biology

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Development of an agroinfiltration-based transient hairpin RNA expression system in pak choi leaves (Brassica rapa ssp. chinensis) for RNA interference against Liriomyza sativae

Li,

Chen,

Gao

et al. 2024

Pesticide Biochemistry and Physiology

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Simultaneous silencing of gut nucleases and a vital target gene by adult dsRNA feeding enhances RNAi efficiency and mortality inCeratitis capitataadults

Volpe,

Mazzucchiello,

Rosati

et al. 2024

Preprint

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Ceratitis capitata, known as Mediterranean fruit fly (Medfly), is a major dipteran pest significantly impacting fruit and vegetable farming. Currently, its control heavily relies mainly on chemical insecticides, which pose health risks and have effects on pollinators. A friendly and species-specific alternative strategy involves providing double-stranded RNA (dsRNA) through feeding to disrupt essential functions in pest insects, which is poorly explored in dipteran species. Previous reports in Orthoptera and Coleoptera species suggested that dsRNA degradation by two specific nucleases in the intestinal lumen is among the major obstacle to feeding-mediated RNAi in insects. In our study, we experimented with three-day adult feeding using a combination of dsRNA molecules that target the expression of theATPasevital gene and two intestinal dsRNA nucleases. These dsRNA molecules were recently tested separately in two Tephritidae species, showing limited effectiveness [1,2]. In contrast, we observed 79% mortality over seven days, which was associated with a decrease in mRNA levels of the three targeted genes. As expected, we also observed a reduction in dsRNA degradation following RNAi against nucleases. This research illustrates the potential of utilizing molecules as pesticides to achieve mortality rates in Medfly adults by targeting crucial genes and intestinal nucleases. Furthermore, it underscores the importance of exploring RNAi-based approaches for pest managementSimple SummaryThe control of insect pest species, mainly belonging to Orthoptera, Hemiptera, and Coleoptera species, can be based on novel emerging species-specific pesticides. These consist of dsRNA molecules delivered by feeding to insect larvae or adults, which suppress vital gene functions by RNA-RNA sequence complementarity and RNA interference. However, fewer studies of dsRNA feeding have been performed in dipteran pest insects. Two studies in Orthoptera and Coleoptera species have shown that suppressing intestinal enzymes degrading external dsRNA can improve insect mortality rates.Ceratitis capitata(Tephritidae), the Mediterranean fruit fly (Medfly), is a major dipteran pest significantly impacting fruit and vegetable farming. Currently, its control heavily relies mainly on chemical insecticides, which pose health risks and have effects on beneficial pollinators. Previous attempts to induce mortality by adult dsRNA-feeding in this and other Tephritidae species, such asBactrocera tryoniandB. dorsalis, showed some effectiveness, but were often limited. We improved this method by simultaneously silencing two intestinal nucleases and a vital gene. We have found a mix of three dsRNAs able to induce much higher mortality (79%) within one week, following only three days of adult feeding.

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RNAi‐mediated silencing of Mediterranean fruit fly (Ceratitis capitata) endogenous genes using orally‐supplied double‐stranded RNAs produced in Escherichia coli

Cited by 4 publications

References 91 publications

RNA Interference in Insects: From a Natural Mechanism of Gene Expression Regulation to a Biotechnological Crop Protection Promise

RNA Interference in Insects: From a Natural Mechanism of Gene Expression Regulation to a Biotechnological Crop Protection Promise

Development of an agroinfiltration-based transient hairpin RNA expression system in pak choi leaves (Brassica rapa ssp. chinensis) for RNA interference against Liriomyza sativae

Simultaneous silencing of gut nucleases and a vital target gene by adult dsRNA feeding enhances RNAi efficiency and mortality inCeratitis capitataadults

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